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Autophagy enhancement in septic liver injury can play a protective role. Nerveless, the mechanism of autophagy-mediated septic liver injury needs further investigation. Our study demonstrated that in septic condition, GLI Family Zinc Finger 2 (GLI2) was elevated, whereas peroxisome-proliferator-activated receptor α (PPARα) was downregulated. Suppressing GLI2 or synovialapoptosis inhibitor 1 (SYVN1) in LPS-exposed cells increased PPARα levels, enhanced cell viability and autophagy, while inhibiting apoptosis. LPS enhanced the GLI2-SYVN1 promoter binding. SYVN1 fostered ubiquitin-mediated degradation of PPARα. IGF2BP3 stabilized GLI2 mRNA by targeting its m6A site. Silencing IGF2BP3 led to decreased GLI2 and SYVN1 but increased PPARα levels, promoting cell survival and autophagy, while repressing apoptosis. This was counteracted by SYVN1 overexpression. In cecal ligation and puncture mice, IGF2BP3, SYVN1, or GLI2 knockdown ameliorated liver damage and augmented autophagy. In summary, IGF2BP3 enhanced GLI2 stability, overexpressed GLI2 subsequent promoted SYVN1 levels by interacting with its promoter, leading to ubiquitinated degradation of PPARα, thereby inhibiting PPARα-mediated autophagy and then exacerbating liver injury in sepsis.
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BACKGROUND: Gliomas advance rapidly and are associated with a poor prognosis. Epithelial-mesenchymal transition (EMT) accelerates the progression of gliomas, exerting a pivotal role in glioma development. Proteasome subunit alpha type-2 (PSMA2) exhibits high expression levels in gliomas. however, its specific involvement in glioma progression and its correlation with EMT remain elusive. This study aims to elucidate the role of PSMA2 in glioma progression and its potential association with EMT. METHODS: Online tools were employed to analyze the expression patterns and survival curves of PSMA2 in gliomas. The relationship between PSMA2 and various characteristics of glioma patients was investigated using data from the TCGA and CGGA databases. In vitro, cell proliferation and migration were assessed through CCK-8, colony formation, and transwell assays. Furthermore, a tumor xenograft model in nude mice was established to evaluate in vivo tumorigenesis. Protein binding to PSMA2 was scrutinized using co-immunoprecipitation MS (co-IP MS). The potential biological functions and molecular pathways associated with PSMA2 were explored through GO analysis and KEGG analysis, and the correlation between PSMA2 and EMT was validated through correlation analysis and Western blot experiments. RESULTS: Bioinformatics analysis revealed a significant upregulation of PSMA2 across various cancers, with particularly heightened expression in gliomas. Moreover, elevated PSMA2 levels were correlated with advanced tumor stages and diminished survival rates among glioma patients. Inhibition of PSMA2 demonstrated a pronounced suppressive effect on glioma cell proliferation, both in vitro and in vivo. Knockdown of PSMA2 also impeded the migratory capacity of glioma cells. GO and KEGG enrichment analyses indicated that PSMA2-binding proteins (identified through Co-IP-MS) were associated with cell adhesion molecule binding and cadherin binding. Western blot results further confirmed the role of PSMA2 in promoting epithelial-mesenchymal transition (EMT) in glioma cells. CONCLUSION: Our study provides evidence supporting the role of PSMA2 as a regulatory factor in EMT and suggests its potential as a prognostic biomarker for glioma progression.
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Glioma , Animales , Humanos , Ratones , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular/genética , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Glioma/patología , Ratones DesnudosRESUMEN
Background: Sepsis is a leading cause of morbidity and mortality in hospitalized patients. Up to now, there are no well-established longitudinal networks from molecular mechanisms to clinical phenotypes in sepsis. Adding to the problem, about one of the five patients presented with diabetes. For this subgroup, management is difficult, and prognosis is difficult to evaluate. Methods: From the three databases, a total of 7,001 patients were enrolled on the basis of sepsis-3 standard and diabetes diagnosis. Input variable selection is based on the result of correlation analysis in a handpicking way, and 53 variables were left. A total of 5,727 records were collected from Medical Information Mart for Intensive Care database and randomly split into a training set and an internal validation set at a ratio of 7:3. Then, logistic regression with lasso regularization, Bayes logistic regression, decision tree, random forest, and XGBoost were conducted to build the predictive model by using training set. Then, the models were tested by the internal validation set. The data from eICU Collaborative Research Database (n = 815) and dtChina critical care database (n = 459) were used to test the model performance as the external validation set. Results: In the internal validation set, the accuracy values of logistic regression with lasso regularization, Bayes logistic regression, decision tree, random forest, and XGBoost were 0.878, 0.883, 0.865, 0.883, and 0.882, respectively. Likewise, in the external validation set 1, lasso regularization = 0.879, Bayes logistic regression = 0.877, decision tree = 0.865, random forest = 0.886, and XGBoost = 0.875. In the external validation set 2, lasso regularization = 0.715, Bayes logistic regression = 0.745, decision tree = 0.763, random forest = 0.760, and XGBoost = 0.699. Conclusion: The top three models for internal validation set were Bayes logistic regression, random forest, and XGBoost, whereas the top three models for external validation set 1 were random forest, logistic regression, and Bayes logistic regression. In addition, the top three models for the external validation set 2 were decision tree, random forest, and Bayes logistic regression. Random forest model performed well with the training and three validation sets. The most important features are age, albumin, and lactate.
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Diabetes Mellitus , Sepsis , Humanos , Mortalidad Hospitalaria , Teorema de Bayes , Aprendizaje Automático , Sepsis/diagnóstico , Ácido LácticoRESUMEN
BACKGROUND: The global volume of gastrointestinal surgery has increased steadily. However, there is still a lack of studies focused on the risk factors for post-gastrointestinal resection surgery patients in the intensive care units. METHODS: Post gastrointestinal resection surgery patient data were collected from the Medical Information Mart for Intensive Care (MIMIC-III) database and divided into training set and validation set, then analyzed by Univariate and multiple logistic regression. RESULTS: 795 patients were finally enrolled in our cohort. Multiple logistic regression showed that age (1.029 [1.006-1.053]), temperature (0.337 [0.207-0.547]), respiratory rate (1.133 [1.053-1.218]), mean arterial pressure (1.204 [1.039-1.396]), lactate (1.288 [1.112-1.493]), BUN (1.025 [1.010-1.040]) and vasopressor use (4.777 [2.499-9.130]) were independent factors associated with in-hospital mortality. Our new predicted nomogram achieved a better accuracy than SOFA score, SAPS-â ¡ score, APACHE-â ¢ score, and Elixhauser score. CONCLUSION: Our nomogram model could well predict in-hospital mortality for post-GI resection surgery patients receiving intensive care.
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Unidades de Cuidados Intensivos , Nomogramas , APACHE , Mortalidad Hospitalaria , Humanos , Pronóstico , Curva ROC , Estudios RetrospectivosRESUMEN
Our study aimed to explore the function of circRNA_0001805 in the pathogenesis of NAFLD and the underlying mechanism. A nanodrug system (GA-RM/GZ/PL) was constructed to overexpress circRNA_0001805 specifically in hepatocytes for the treatment of NAFLD. Fat droplet accumulation in cultured cells and mouse hepatic tissues was detected using Oil Red O or H&E staining. The relative expression of circRNAs, genes associated with lipogenesis was quantified by qRT-PCR. Interactions between circRNA_0001805 and miR-106a-5p/miR-320a, between miR-106a-5p/miR-320a and ABCA1/CPT1 were confirmed by dual-luciferase reporter assay. A novel metalorganic framework nanocarrier (GZ) was prepared from glycyrrhizic acid and zinc ions (Zn2+), and this nanocarrier was loaded with the circRNA_0001805 plasmid to construct a nanocore (GZ/PL). Then, this GZ/PL was coated with a galactose-modified RBC membrane (GA-RM) to generate GA-RM/GZ/PL. CircRNA_0001805 expression was downregulated in FFA-challenged primary hepatocytes, HFD-fed mice and NAFLD patients. Overexpressed circRNA_0001805 attenuated NAFLD development by suppressing lipid metabolism disorder and inflammation. CircRNA_0001805 targeted miR-106a-5p/miR-320a, which served as an upstream inhibitor of ABCA1/CPT1 and collaboratively regulated NAFLD progression. GA-RM/GZ/PL targeted hepatocytes, overexpressed circRNA_0001805, released glycyrrhizic acid to reduce the accumulation of lipids in the liver and played a synergistic role against NAFLD-induced lipid metabolism disorder.
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Transportador 1 de Casete de Unión a ATP , Carnitina O-Palmitoiltransferasa , Nanoestructuras/química , Enfermedad del Hígado Graso no Alcohólico/metabolismo , ARN Circular , Transportador 1 de Casete de Unión a ATP/genética , Transportador 1 de Casete de Unión a ATP/metabolismo , Animales , Carnitina O-Palmitoiltransferasa/genética , Carnitina O-Palmitoiltransferasa/metabolismo , Humanos , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Nanomedicina/métodos , ARN Circular/genética , ARN Circular/metabolismo , ARN Circular/farmacologíaRESUMEN
OBJECTIVES: Non-alcoholic fatty liver disease has seriously affected people's health. Recent studies have found that N6-methyladenosine (m6A) methylation is involved in the lipid metabolism process of the body, but the study on the level of m6A modification in NAFLD is still not available. This study aims to explore the changes in the level of RNA m6A methylation modification in NAFLD liver tissues, and to provide experimental and theoretical basis for in-depth study on the role of RNA m6A methylation in the occurrence and development of NAFLD. METHODS: Changes in the m6A level in NAFLD liver tissues were measured by liquid chromatography-mass spectrometry (LC-MS). Total RNA was extracted from liver tissues of NAFLD patients or normal control individuals and subjected to methylated RNA immunoprecipitation (MeRIP) with microarray analysis (including 44 122 mRNAs and 12 496 lncRNAs) to determine the changes in m6A modification levels across the entire transcriptome. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to annotate the differentially modified mRNAs. Finally, 4 mRNAs and 4 lncRNAs were randomly selected to verify the microarray results by MeRIP and real-time transcription polymerase chain reaction. RESULTS: A total of 176 mRNAs and 44 lncRNAs were found to be differentially m6A-modified in the NAFLD group compared with the control group. Among them, 15 mRNAs and 7 lncRNAs were hypermethylated in NAFLD, while 161 mRNAs and 37 lncRNAs were hypomethylated in NAFLD. GO and pathway analysis showed that the differentially modified mRNAs were enriched mainly in biological processes such as carboxylic acid metabolism and transcriptional regulation. CONCLUSIONS: The m6A modification profile is changed in NAFLD liver tissues compared with normal liver tissues, which may functionally impact the pathophysiological progress in NAFLD.
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Enfermedad del Hígado Graso no Alcohólico , ARN Largo no Codificante , Humanos , Enfermedad del Hígado Graso no Alcohólico/genética , ARN Largo no Codificante/genética , ARN Mensajero/genética , TranscriptomaRESUMEN
Fibrotic tissue remodelling in nonalcoholic fatty liver disease (NAFLD) will probably emerge as the leading cause of end-stage liver disease in the coming decades, but the ability to diagnose liver fibrosis in NAFLD patients noninvasively is limited. The abnormal expression of tRNA-derived small RNA (tsRNA) in plasma provides a novel idea for noninvasive diagnosis of various diseases, however, the relationship between tsRNAs and NAFLD is still unknown. Here, we took advantage of small RNA-Seq technology to profile tsRNAs in NAFLD patients and found the ubiquitous presence of hepatic tsRNAs secreted into circulating blood. Verification in a cohort of 114 patients with NAFLD and 42 patients without NAFLD revealed that three tsRNAs (tRF-Val-CAC-005, tiRNA-His-GTG-001, and tRF-Ala-CGC-006) were significantly elevated in the plasma of NAFLD patients, and the expression level are associated with NAFLD activity score (calculated from 0 to 8) and fibrosis stage (scored from 0 to 4). In mouse models, we further found that increased plasma levels of these three tsRNAs were positively correlated with the degree of liver fibrosis. Our study potentially identifies a new class of NAFLD biomarkers and reveal the possible existence of tsRNAs in the blood that can be used to predict fibrogenesis risk in patients diagnosed with NAFLD.
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Cirrosis Hepática/sangre , Enfermedad del Hígado Graso no Alcohólico/sangre , ARN de Transferencia/sangre , Adulto , Anciano , Animales , Secuencia de Bases , Biomarcadores/sangre , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Perfilación de la Expresión Génica , Humanos , Hígado/metabolismo , Masculino , Ratones Endogámicos BALB C , Persona de Mediana Edad , ARN de Transferencia/química , ARN de Transferencia/genética , Regulación hacia Arriba/genética , Adulto JovenRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is regarded as a threat to public health; however, the pathologic mechanism of NAFLD is not fully understood. We attempted to identify abnormally expressed long noncoding RNA (lncRNAs) and messenger RNA that may affect the occurrence and development of NAFLD in this study. The expression of differentially expressed lncRNAs in NAFLD was determined in oleic acid (OA)-treated L02 cells, and the functions of CCAT1 in lipid droplet formation were evaluated in vitro. Differentially expressed genes (DEGs) were analyzed by microarray analysis, and DEGs related to CCTA1 were selected and verified by weighted correlation network analysis. The dynamic effects of LXRα and CCTA1 on lipid droplet formation and predicted binding was examined. The binding between miR-631 and CCAT1 and LXRα was verified. The dynamic effects of miR-613 inhibition and CCTA1 silencing on lipid droplet formation were examined. The expression and correlations of miR-631, CCAT1, and LXRα were determined in tissue samples. As the results show, CCAT1 was induced by OA and upregulated in NAFLD clinical samples. CCAT1 silencing significantly suppressed lipid droplet accumulation in vitro. LXRα was positively correlated with CCAT1. By inhibiting miR-613, CCAT1 increased the transcription of LXRα and promoted LXRα expression. The expression of LXRα was significantly increased in NAFLD tissues and was positively correlated with CCAT1. In conclusion, CCAT1 increases LXRα transcription by serving as a competing endogenous RNA for miR-613 in an LXRE-dependent manner, thereby promoting lipid droplet formation and NAFLD. CCAT1 and LXRα might be potent targets for NAFLD treatment.
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Receptores X del Hígado/genética , MicroARNs/genética , Enfermedad del Hígado Graso no Alcohólico/genética , ARN Largo no Codificante/genética , Línea Celular Tumoral , Proliferación Celular/genética , Regulación de la Expresión Génica/genética , Humanos , Gotas Lipídicas/metabolismo , Gotas Lipídicas/patología , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Transcripción Genética/genéticaRESUMEN
BACKGROUND: Hepatic lipogenesis dysregulation is essential for the development of non-alcoholic fatty liver disease (NAFLD). Emerging evidence indicates the importance of the involvement of long non-coding RNAs (LncRNAs) in lipogenesis. However, the specific mechanism underlying this process is not clear. OBJECTIVE: This study aimed to investigate the functional implication of LncRNA MEG3 (MEG3) in fatty degeneration of hepatocytes and in the pathogenesis of NAFLD. METHODS: The expression of MEG3 was analysed in in vitro and in vivo models of NAFLD, which were established by free fatty acid (FFA)-challenged HepG2 cells and high-fat diet-fed mice, respectively. Endogenous MEG3 was over-expressed by a specific pcDNA3.1-MEG3 to evaluate the regulatory function of MEG3 on triglyceride (TG)- and lipogenesis-related genes. Bioinformatic analysis was used to predict the target genes and binding sites, and the targeted regulatory relationship was verified with a dual luciferase assay. Finally, the possible pathway that regulates MEG3 was also evaluated. RESULTS: We found that the downregulation of MEG3 in vitro and in vivo models of NAFLD was negatively correlated with lipogenesis-related genes and that overexpression of MEG3 reversed FFA-induced lipid accumulation in HepG2 cells. miR-21 was upregulated in the FFA-challenged HepG2 cells and was physically associated with MEG3 in the process of lipogenesis. Our mechanistic studies demonstrated that MEG3 competitively binds to miR-21 with LRP6, followed by the inhibition of the mTOR pathway, which induces intracellular lipid accumulation. CONCLUSION: Our data are the first to document the working model of MEG3 functions as a potential hepatocyte lipid degeneration suppressor. MEG3 helps to alleviate lipid over-deposition, probably by binding to miR-21 to regulate the expression of LRP6. Our results suggest the potency of MEG3 as a biomarker for NAFLD and as a therapeutic target for treatment.
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Lipogénesis , Hígado/metabolismo , Proteína-6 Relacionada a Receptor de Lipoproteína de Baja Densidad/metabolismo , MicroARNs/metabolismo , ARN Largo no Codificante/fisiología , Animales , Unión Competitiva , Células Hep G2 , Humanos , Lipogénesis/efectos de los fármacos , Lipogénesis/genética , Ratones , Enfermedad del Hígado Graso no Alcohólico/diagnóstico , Enfermedad del Hígado Graso no Alcohólico/etiología , Enfermedad del Hígado Graso no Alcohólico/metabolismo , ARN Largo no Codificante/farmacologíaRESUMEN
BACKGROUND: Acute lung injury (ALI) is a life-threatening lung disease where alveolar macrophages (AMs) play a central role both in the early phase to initiate inflammatory responses and in the late phase to promote tissue repair. In this study, we examined whether BML-111, a lipoxin A4 receptor agonist, could alter the phenotypes of AM and thus present prophylactic benefits for ALI. METHODS: In vitro, isolated AMs were treated with lipopolysaccharide (LPS) to induce ALI. In response to BML-111 pre-treatment, apoptosis and autophagy of AMs were examined by flow cytometry, and by measuring biomarkers for each process. The potential involvement of MAPK1 and mTOR signaling pathway was analyzed. In vivo, an LPS-induced septic ALI model was established in rats and the preventative significance of BML-111 was assessed. On the cellular and molecular levels, the pro-inflammatory cytokines TNF-α and IL-6 from bronchoalveolar lavage were measured by ELISA, and the autophagy in AMs examined using Western blot. RESULTS: BML-111 inhibited apoptosis and induced autophagy of AMs in response to ALI inducer, LPS. The enhancement of autophagy was mediated through the suppression of MAPK1 and MAPK8 signaling, but independent of mTOR signaling. In vivo, BML-111 pre-treatment significantly alleviated LPS-induced ALI, which was associated with the reduction of apoptosis, the dampened production of pro-inflammatory cytokines in the lung tissue, as well as the increase of autophagy of AMs. CONCLUSIONS: This study reveals the prophylactic significance of BML-111 in ALI and the underlying mechanism: by targeting the MAPK signaling but not mTOR pathway, BML-111 stimulates autophagy in AMs, attenuates the LPS-induced cell apoptosis, and promotes the resolution of ALI.
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Lesión Pulmonar Aguda/enzimología , Autofagia/fisiología , Ácidos Heptanoicos/uso terapéutico , Sistema de Señalización de MAP Quinasas/fisiología , Macrófagos Alveolares/enzimología , Receptores de Lipoxina/agonistas , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/prevención & control , Animales , Autofagia/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Activación Enzimática/fisiología , Ácidos Heptanoicos/farmacología , Lipopolisacáridos/toxicidad , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Macrófagos Alveolares/efectos de los fármacos , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To analyze the characteristics of long non-coding RNA (lncRNA) expression in non-alcoholic fatty liver disease (NAFLD).â© Methods: lncRNA-mRNA microarray was conducted on the liver tissue samples from 10 patients with simple gallbladder stone (5 NAFLD liver samples and 5 normal liver samples), and the differentially expressed lncRNA was analyzed by bioinformatics technology.â© Results: Compared with the normal liver samples, there were abnormal expression of 1 735 lncRNAs and 1 485 mRNAs in NAFLD liver samples. Among them, 535 lncRNAs and 760 mRNAs were up-regulated, 1 200 lncRNAs and 725 mRNAs were down-regulated.â© Conclusion: Compared with normal liver, the expression of lncRNA in NAFLD tissues is obviously abnormal. These lncRNAs may play an important role in the occurrence and development of NAFLD.
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Regulación de la Expresión Génica , Enfermedad del Hígado Graso no Alcohólico/fisiopatología , ARN Largo no Codificante/genética , Enfermedades de la Vesícula Biliar/fisiopatología , Perfilación de la Expresión Génica , Humanos , ARN Mensajero/genéticaRESUMEN
Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder poising burgeoning health problem to humans. Recent studies have shown that long non-coding RNA (lncRNA) plays critical roles in a myriad of biological processes and human diseases. Since the roles of lncRNA in NAFLD remain unknown, they were investigated in the study. Microarray expression profiling of mRNAs and lncRNAs was conducted using RNA extracted from patients with and without NAFLD. One thousand seven hundred thirty-five lncRNAs and 1485 mRNA were found differentially expressed in NAFLD samples compared with those in control samples. Among them 535 and 1,200 lncRNAs were upregulated and downregulated in NAFLD, respectively; 760 and 725 mRNAs were upregulated and downregulated in NAFLD, respectively. Moreover, seven lncRNAs and seven mRNAs that were highly up- or downregulated in NAFLD samples were validated by quantitative real-time polymer chain reactions. Kyoto Encyclopedia of Genes and Genomes pathway analysis and Gene Ontology analysis for the differentially expressed mRNAs showed that these RNAs are involved in various metabolic processes, cellular components, and molecular functions. Our findings indicate that the expression profiles of lncRNAs have changed in NAFLD as compared with normal liver, and the identified regulated RNAs may provide novel insight into the molecular mechanisms underlying the disease and potential novel diagnostic or therapeutic targets for NAFLD.
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Enfermedad del Hígado Graso no Alcohólico/metabolismo , ARN Largo no Codificante/metabolismo , Adulto , Biomarcadores/metabolismo , Estudios de Casos y Controles , Femenino , Perfilación de la Expresión Génica , Ontología de Genes , Genoma Humano , Humanos , Hígado/metabolismo , Masculino , Persona de Mediana Edad , ARN Largo no Codificante/genética , TranscriptomaRESUMEN
Traumatic hemorrhagic shock (HS) is a severe outcome of traumatic injury that accounts for numerous traumatic deaths. In the process of traumatic HS, both hemorrhage and trauma can trigger a complex cascade of posttraumatic events that are related to inflammatory and immune responses, which may lead to multiple organ injury or even death. From a mechanistic perspective, systemic inflammation and organ injury are involved coagulation, the complement system, impaired microcirculation and inflammatory signaling pathways. In this review, we discuss the systemic inflammation and multiple organ injury in post-traumatic HS.
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Insuficiencia Multiorgánica/complicaciones , Choque Hemorrágico/patología , Animales , Modelos Animales de Enfermedad , Humanos , Inmunidad Innata , Inflamación , Hepatopatías/complicaciones , Hepatopatías/patología , Ratones , Insuficiencia Multiorgánica/patología , Ratas , Choque Hemorrágico/complicaciones , Choque Hemorrágico/inmunologíaRESUMEN
5(S),6(R)-7-trihydroxymethyl heptanoate (BML-111) is an lipoxin A4 receptor agonist, which modulates the immune response and attenuates hemorrhagic shock-induced acute lung injury. However, the role of BML-111 in sepsis and in the intestinal mucosal barrier are not well understood. Therefore, the present study was designed to investigate the effect of BML-111 on the intestinal mucosal barrier in a rat model of sepsis. Furthermore, the molecular mechanism of action of BML-111 was evaluated. The cecal ligation and puncture-induced rat model of sepsis was constructed, and BML-111 was administered at three different doses. The results revealed that BML-111 suppressed the elevation of the pro-inflammatory cytokines tumor necrosis factor-α and interleukin-6, while enhancing the elevation of the anti-inflammatory cytokine transforming growth factor-ß in the intestine. In addition, BML-111 significantly upregulated rat defensin-5 mRNA expression levels and downregulated the induction of cell apoptosis as well as caspase-3 activity in the intestine. All these results demonstrated that BML-111 exerted protective effects on the intestinal mucosal barrier in sepsis. Further, it was indicated that alterations in the expression of toll-like receptor (TLR)2 and TLR4 may be one of the molecular mechanisms underlying the protective effect of BML-111. The present study therefore suggested that BML-111 may be a novel therapeutic agent for sepsis.
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Antiinflamatorios no Esteroideos/farmacología , Defensinas/agonistas , Ácidos Heptanoicos/farmacología , Mucosa Intestinal/efectos de los fármacos , Sepsis/tratamiento farmacológico , Animales , Apoptosis , Caspasa 3/genética , Caspasa 3/inmunología , Defensinas/genética , Defensinas/inmunología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Humanos , Inmunidad Mucosa/efectos de los fármacos , Inyecciones Intraperitoneales , Interleucina-6/genética , Interleucina-6/inmunología , Mucosa Intestinal/inmunología , Mucosa Intestinal/patología , Masculino , Ratas , Ratas Sprague-Dawley , Sepsis/genética , Sepsis/inmunología , Sepsis/patología , Transducción de Señal , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunología , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/inmunología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Non-alcoholic fatty liver disease (NAFLD) has emerged as a public health issue with a prevalence of 15-30% in Western populations and 6-25% in Asian populations. Certain studies have revealed the alteration of microRNA (miRNA or miR) profiles in NAFLD and it has been suggested that miR-21 is associated with NAFLD. In the present study, we measured the serum levels of miR-21 in patients with NAFLD and also performed in vitro experiments using a cellular model of NAFLD to further investigate the effects of miR-21 on triglyceride and cholesterol metabolism. Furthermore, a novel target through which miR-21 exerts its effects on NAFLD was identified. The results revealed that the serum levels of miR-21 were lower in patients with NAFLD compared with the healthy controls. In addition, 3-hydroxy-3-methylglutaryl-co-enzyme A reductase (HMGCR) expression was increased in the serum of patients with NAFLD both at the mRNA and protein level. To mimic the NAFLD condition in vitro, HepG2 cells were treated with palmitic acid (PA) and oleic acid (OA). Consistent with the results obtained in the in vivo experiments, the expression levels of miR-21 were decreased and those of HMGCR were increased in the in vitro model of NAFLD. Luciferase reporter assay revealed that HMGCR was a direct target of miR-21 and that miR-21 exerted an effect on both HMGCR transcript degradation and protein translation. Furthermore, the results from the in vitro experiments revealed that miR-21 decreased the levels of triglycerides (TG), free cholesterol (FC) and total cholesterol (TC) in the PA/OA-treated HepG2 cells and that this effect was attenuated by HMGCR overexpression. Taken together, to the best of our knowledge, the present study is the first to report that miR-21 regulates triglyceride and cholesterol metabolism in an in vitro model of NAFLD, and that this effect is achieved by the inhibition of HMGCR expression. We speculate that miR-21 may be a useful biomarker for the diagnosis and treatment of NAFLD.
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Colesterol/metabolismo , Regulación de la Expresión Génica , Hidroximetilglutaril-CoA Reductasas/genética , MicroARNs/genética , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Triglicéridos/metabolismo , Regiones no Traducidas 3' , Estudios de Casos y Controles , Células Hep G2 , Humanos , Hidroximetilglutaril-CoA Reductasas/química , Hidroximetilglutaril-CoA Reductasas/metabolismo , MicroARNs/química , Interferencia de ARN , ARN Mensajero/química , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
To explore protective mechanism of Panax notoginseng saponins (PNS) on rat hemorrhagic shock model in recovery stage. 72 Wistar rats were selected and divided into control group, model group and PNS group with 24 rats in each group. 200 mg/kg PNS was injected intravenously at 60 min of hemorrhagic shock stage in PNS groups. Changes of endotoxin, MPO, IL-6, SOD, MDA and TNF α were observed at 30 and 120 min of recovery stage by ELISA; water content of lung and intestine was detected; HE staining was applied to observe morphological change of intestinal mucosa, kidney, liver and lung; western blot was used to detect intercellular adhesion molecule-1 (ICAM-1) level in lung tissue and intestine tissue. At 30 min and 120 min of recovery stage, MDA, MPO, endotoxin, TNF α and IL-6 levels significantly increased in model group compared with control group, however SOD level significantly decreased, the difference was statistically significant (P < 0.05); PNS dose-dependently decreased MDA, MPO, endotoxin, TNF α and IL-6 levels, and increased SOD level, which was statistically significant (P < 0.05); In results of water content detection, water content in lung tissue and intestine tissue was significantly higher than in control group, however, after being treated with PNS, the water content was significantly decreased; HE staining showed the morphologic change of lung tissue cells; Western blot showed that in lung tissue and intestine tissue, ICAM-1 level in model group was significantly higher than in control group, and it was lower in PNS group than in model group. PNS can increase SOD activity, decrease levels of MDA, endotoxin and MPO, decrease expression of TNF α and IL-6, and decrease water content in lung tissue and intestine tissue. Thus, PNS is protective to rat hemorrhagic shock model by anti oxidative stress and anti-inflammatory pathways, and ICAM-1 may play an important role in the mechanism.
Asunto(s)
Sustancias Protectoras/uso terapéutico , Saponinas/uso terapéutico , Choque Hemorrágico/tratamiento farmacológico , Animales , Modelos Animales de Enfermedad , Endotoxinas/sangre , Ensayo de Inmunoadsorción Enzimática , Molécula 1 de Adhesión Intercelular/metabolismo , Interleucina-6/sangre , Pulmón/metabolismo , Pulmón/patología , Masculino , Malondialdehído/sangre , Panax notoginseng/química , Panax notoginseng/metabolismo , Peroxidasa/sangre , Ratas , Ratas Wistar , Superóxido Dismutasa/sangre , Factor de Necrosis Tumoral alfa/sangreRESUMEN
OBJECTIVE: To explore the molecular mechanism of fibroblast growth factor 8b (FGF8b) in promoting epithelial-mesenchymal transition in prostate cancer DU145 cells. METHODS: Cells were selected in three groups as follows: a block control group (DU145 cells), a negative control group [DU145 cells transfected with empty plasmid (pcDNA3.1/DU145)], and an experimental group [DU145 cells transfected with FGF8b (FGF8b/DU145)]. The activity of extracellular regulated protein kinases1/2( ERK1/2) pathway was detected by western-blot in the three groups. The FGF8b-DU145 cells and DU145 cells were cultured with PD98059 (an ERK kinase inhibitor) to observe microscopically the morphology changes within the cells. The experimental samples were also divided into four groups: FGF8b/DU145 cells cultured with 2% FBS (Group A); FGF8b/DU145 cells cultured with 2% FBS+PD98059 (50 µmol/L) (Group B); DU145 cells cultured with 2% FBS (Group C); DU145 cells cultured with FBS+PD98059 (50 µmol/L) (Group D). The expression of epithelial- mesenchymal transition (EMT) markers (E-cadherin, vimentin) were detected by western-blot analysis and the cell's mobility were detected by the Transwell chamber. RESULTS: The activity of ERK1/2 in the experimental group was significantly higher than that in the other two control groups; when ERK kinase inhibitor PD98059 was added to FGF8b/ DU145 cells, the expression of epithelial marker E-cadherin protein was significantly increased in group B compared with that in the group A (P<0.05). The expression of mesenchymal marker vimentin protein was significantly reduced in group B compared with that in group A (P<0.05). The cell migration assay suggested that cell migration was markedly decreased in group B (P<0.05) compared with that in group A. CONCLUSION: EMT in prostate cancer induced by FGF8b can be mediated by ERK kinase pathway, in which mitogen-activated/extraceluer signal regulated kinase 1 (MEK1) may be a key factor. MEK1 could be an effective target in regulating the invasion and migration of prostate cancer.
Asunto(s)
Transición Epitelial-Mesenquimal/genética , Factor 8 de Crecimiento de Fibroblastos/metabolismo , Neoplasias de la Próstata/patología , Factor 8 de Crecimiento de Fibroblastos/genética , Flavonoides/farmacología , Humanos , MAP Quinasa Quinasa 1/metabolismo , Sistema de Señalización de MAP Quinasas/fisiología , Masculino , Invasividad Neoplásica , Metástasis de la Neoplasia , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Transfección , Células Tumorales CultivadasRESUMEN
OBJECTIVE: Overexpression of elongation factor-1α (EF-1α) has been demonstrated to be related to increased cell proliferation, oncogenic transformation and delayed cell senescence. The purpose of this study was to determine whether EF-1α expression affects the progression of prostate cancer (PCa), and whether it can be used as a prognostic marker for PCa. MATERIAL AND METHODS: EF-1α was evaluated by immunostaining in paraffin-embedded specimens of prostates obtained from 80 patients with PCa. Correlations of EF-1α with patients' ages, Gleason scores, American Joint Committee on Cancer (AJCC) stages, International Union Against Cancer (UICC) stages, preoperative prostate-specific antigen (PSA) concentrations and PSA failure were evaluated. Survival in all patients was analysed to evaluate the influence of EF-1α expression in cancer progression using Kaplan-Meier and multivariate Cox regression analysis. RESULTS: The positive expression rate of EF-1α in PCa tissues [64/80 (80.0%)] was significantly higher than that in normal prostate tissues [1/20 (5.0%)] (p < 0.001). Increased immunostaining of EF-1α was a significant predictor of distant metastasis-free survival [hazard ratio (HR) 0.386, 95% confidence interval (CI) 0.032-2.519, p = 0.003] and overall survival (HR 0.305, 95% CI 0.091-0.872, p = 0.005). In multivariate analysis including competing biological variables, EF-1α expression was still significantly linked to distant metastasis-free survival (HR 0.216, 95% CI 0.042-0.876, p = 0.015) and overall survival (HR 0.395, 95% CI 0.116-0.798, p = 0.008). CONCLUSION: These findings provide convincing evidence for the first time that EF-1α correlates closely with the survival of patients with PCa and may be a novel prognostic marker.